2-脱氧葡萄糖对乳腺癌MCF7/ErbB2细胞增殖迁移和侵袭的影响

Effect of 2-DG on proliferation migration and invasion of breast cancer cell line MCF7/ErbB2

目的探讨糖酵解抑制剂2-脱氧葡萄糖(2-Deoxy-D-glucose,2-DG)对乳腺癌MCF7/ErbB2细胞增殖、迁移和侵袭的影响及其分子机制。方法用不同浓度的2-DG(0,0.5,1,2,4,8,16,32mM)处理MCF7/ErbB2细胞48h后,用MTS试剂盒测定细胞的增殖;用Transwell实验检测不同浓度2-DG(0,0.5,1,2mM)对MCF7/ErbB2细胞的迁移和侵袭的影响;用不同浓度2-DG(0,0.5,1,2mM)处理MCF7/ErbB2细胞48h后,用葡萄糖和乳酸检测试剂盒分别测定培养基中的葡萄糖浓度和乳酸浓度,并计算细胞对葡萄糖的摄取量和乳酸生成量;用1mM 2-DG处理MCF7/ErbB2细胞0、4、8、12、24h后,用Western blot检测己糖激酶II(hexokinase II,HKII)蛋白表达水平。结果与2-DG未处理组比较,2-DG(0.5~32mM)能抑制MCF7/ErbB2细胞的增殖;2-DG(0.5,1,2mM)能显著抑制MCF7/ErbB2细胞的迁移和侵袭(P<0.001),同时显著降低葡萄糖摄取量和乳酸生成量(P<0.001),而且上述抑制作用均表现出对2-DG剂量的依赖性;1mM 2-DG显著下调MCF7/ErbB2细胞中HKII的蛋白水平。结论 2-DG通过下调HKII蛋白表达降低糖酵解水平,从而抑制乳腺癌MCF7/ErbB2细胞的增殖、迁移和侵袭。

Objective To investigate the effect of glycolysis inhibitor 2-DG on proliferation,migration and invasion of ErbB2 overexpressing breast cancer cell line MCF7/ErbB2 and its molecular mechanism.Methods MCF7/ErbB2 cells were treated with 2-DG(0,0.5,1,2,4,8,16,32 mM)for 48 hours.Cell proliferation was detected by MTS assay.MCF7/ErbB2 cells were treated with different concentration of 2-DG(0,0.5,1,2 mM)for 24 hours.Cell migration and invasion was detected by Transwell assays.The glucose and lactate concentration in the culture media were determined by glucose and lactate assay kit,respectively,when MCF7/ErbB2 cells were treated with 2-DG(0,0.5,1,2 mM)for 48 hours,and then glucose uptake and lactate production by the cells were calculated.Finally,the cells were treated with 2-1 mM DG for different times(0,4,8,12,24 hours)to detect hexokinase II(HKII)protein levels by Western blot.Results Compared with untreated group,2-DG(0.5-32 mM)significantly decreased the proliferation of MCF7/ErbB2,2-DG(0.5,1,2 mM)also inhibits the migration,invasion,glucose uptake and lactate production of MCF7/ErbB2 cells(P<0.001).Inhibition of cell proliferation,migration,invasion and glycolysis was 2-DG dose-dependent.Moreover,1 mM 2-DG downregulated HKII protein levels in MCF7/ErbB2 cells significantly.Conclusion In breast cancer cell line MCF7/ErbB2,2-DG inhibits glycolysis by downregulating HKII protein levels leading to inhibition of proliferation,migration and invasion.